WO2021090720A1 - Dispositif de mesure optique et structure de lentille - Google Patents

Dispositif de mesure optique et structure de lentille Download PDF

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Publication number
WO2021090720A1
WO2021090720A1 PCT/JP2020/040090 JP2020040090W WO2021090720A1 WO 2021090720 A1 WO2021090720 A1 WO 2021090720A1 JP 2020040090 W JP2020040090 W JP 2020040090W WO 2021090720 A1 WO2021090720 A1 WO 2021090720A1
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WIPO (PCT)
Prior art keywords
lens
excitation light
light
lenses
positive
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Application number
PCT/JP2020/040090
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English (en)
Japanese (ja)
Inventor
聡史 長江
晃二 喜田
隆史 加藤
Original Assignee
ソニー株式会社
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Publication date
Application filed by ソニー株式会社 filed Critical ソニー株式会社
Priority to JP2021554896A priority Critical patent/JPWO2021090720A1/ja
Priority to US17/755,387 priority patent/US20220404262A1/en
Publication of WO2021090720A1 publication Critical patent/WO2021090720A1/fr

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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/02—Objectives
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • G01N15/1434—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • G01N15/1456—Electro-optical investigation, e.g. flow cytometers without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
    • G01N15/1459—Electro-optical investigation, e.g. flow cytometers without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals the analysis being performed on a sample stream
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47—Scattering, i.e. diffuse reflection
    • G01N21/49—Scattering, i.e. diffuse reflection within a body or fluid
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/16—Microscopes adapted for ultraviolet illumination ; Fluorescence microscopes
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B7/00—Mountings, adjusting means, or light-tight connections, for optical elements
    • G02B7/02—Mountings, adjusting means, or light-tight connections, for optical elements for lenses
    • G02B7/021—Mountings, adjusting means, or light-tight connections, for optical elements for lenses for more than one lens
    • G01N15/149—
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1006—Investigating individual particles for cytology
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B17/00—Systems with reflecting surfaces, with or without refracting elements
    • G02B17/02—Catoptric systems, e.g. image erecting and reversing system
    • G02B17/026—Catoptric systems, e.g. image erecting and reversing system having static image erecting or reversing properties only
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/10—Beam splitting or combining systems
    • G02B27/1006—Beam splitting or combining systems for splitting or combining different wavelengths
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/10—Beam splitting or combining systems
    • G02B27/14—Beam splitting or combining systems operating by reflection only
    • G02B27/145—Beam splitting or combining systems operating by reflection only having sequential partially reflecting surfaces

Definitions

  • the present disclosure relates to an optical measuring device and a lens structure.
  • a flow cytometer irradiates particles flowing through a flow path formed in a flow cell or microchip with light, detects fluorescence or scattered light emitted from each particle, and executes analysis or the like. It is a device for performing optical measurement using cytometry.
  • Flow cytometers include analyzers for the purpose of sample analysis and sorters that have the function of analyzing samples and separating and collecting only particles with specific characteristics based on the analysis results. Further, a sorter having a function of using cells as a sample and separating and collecting cells based on the analysis result is also called a "cell sorter”.
  • An objective lens used in a general optical measuring device for fluorescence observation or the like is a lens structure formed by combining a plurality of lenses, and an adhesive is used for assembling the objective lens.
  • the optical characteristics of the objective lens deteriorate due to burning of the adhesive due to the strong laser beam, or burning of the outgas emitted from the adhesive and adhering to the lens surface due to the excitation light. There was a problem that it could end up.
  • the present disclosure proposes an optical measuring device and a lens structure capable of suppressing deterioration of optical characteristics.
  • the optical measuring device of one embodiment according to the present disclosure has an excitation light source that emits excitation light having a wavelength of at least 450 nanometers or less, and a lens structure that concentrates the excitation light at a predetermined position.
  • a body a fluorescence detection system that detects fluorescence emitted from the particles by exciting the particles existing at the predetermined positions by the excitation light, and the particles in which the excitation light exists at the predetermined positions.
  • the lens structure includes a plurality of lenses arranged along the optical axis of the excitation light and a lens holding the plurality of lenses, which comprises a scattered light detection system for detecting scattered light generated by being scattered.
  • a frame is provided, and at least one of the plurality of lenses is in contact with a lens adjacent to the lens to determine a position in the lens frame.
  • FIG. 1 It is sectional drawing which shows the schematic structural example of the imaging lens which concerns on one Embodiment of this disclosure. It is a figure which shows an example of the longitudinal aberration of the optical system which combined the objective lens and the imaging lens which concerns on 1st specific example (spherical aberration). It is a figure which shows an example of the longitudinal aberration of the optical system which combined the objective lens and the imaging lens which concerns on 1st specific example (astigmatism). It is a figure which shows an example of the longitudinal aberration of the optical system which combined the objective lens and the imaging lens which concerns on 1st specific example (distortion aberration).
  • a cell analyzer is exemplified as the optical measuring device.
  • the cell analyzer according to the present embodiment may be, for example, a cell sorter type flow cytometer (hereinafter, simply referred to as a cell sorter).
  • the microchip method is exemplified as a method of supplying fine particles to an observation point (hereinafter referred to as a spot) on the flow path, but the method is not limited to this, and for example, a droplet method or a cuvette method is used. , And various methods such as a flow cell method can be adopted. Further, the technique according to the present disclosure is not limited to the cell sorter, and measures fine particles passing through a spot set on the flow path, such as an analyzer-type flow cytometer and a microscope for acquiring an image of fine particles on the flow path. It can be applied to various optical measuring devices.
  • FIG. 1 is a schematic diagram showing a schematic configuration example of the optical system in the cell analyzer according to the present embodiment.
  • the cell analyzer 1 includes, for example, one or more (three in this example) excitation light sources 101 to 103, a total reflection mirror 111, a dichroic mirror 112 and 113, and a perforated mirror 114.
  • the total reflection mirror 111, the dichroic mirrors 112 and 113, the perforated mirror 114, and the dichroic mirror 115 are waveguide optics that guide the excitation lights L1 to L3 emitted from the excitation light sources 101 to 103 on a predetermined optical path. Construct a system.
  • the dichroic mirror 115 has fluorescence (for example, fluorescence L14) and scattered light (for example, back scattering) among the light emitted in a predetermined direction (for example, rearward) from the spot 123a set on the flow path in the microchip 120.
  • a separation optical system that separates the light L12) is formed.
  • the perforated mirror 114 uses an optical path different from the predetermined optical path (for example, an optical path toward the rear scattered light detection system 130 described later) for the scattered light (for example, rear scattered light L12) separated by the separation optical system. It constitutes a reflection optical system that reflects light to.
  • the objective lens 116 constitutes a condensing optical system that focuses the excitation lights L1 to L3 propagating on the predetermined optical path onto the spot 123a set on the flow path in the microchip 120.
  • the number of spots 123a is not limited to one, that is, the excitation lights L1 to L3 may be focused on different spots. Further, the focusing positions of the excitation lights L1 to L3 do not have to coincide with the spot 123a, and may be deviated.
  • the excitation light sources 101 to 103 that emit excitation lights L1 to L3 having different wavelengths are provided.
  • a laser light source that emits coherent light may be used.
  • the excitation light source 102 may be a DPSS laser (Diode Pumped Solid State Laser: semiconductor laser excited solid-state laser) that irradiates a blue laser beam (peak wavelength: 488 nm (nanometer), output: 20 mW).
  • DPSS laser Diode Pumped Solid State Laser: semiconductor laser excited solid-state laser
  • the excitation light source 101 may be a laser diode that irradiates a red laser beam (peak wavelength: 637 nm, output: 20 mW), and similarly, the excitation light source 103 may be a near-ultraviolet laser beam (peak wavelength: 405 nm, output). : It may be a laser diode that irradiates 8 mW). Further, the excitation lights L1 to L3 emitted by the excitation light sources 101 to 103 may be pulsed light.
  • the total reflection mirror 111 may be, for example, a total reflection mirror that reflects the excitation light L1 emitted from the excitation light source 101 in a predetermined direction.
  • the dichroic mirror 112 is an optical element for aligning or paralleling the optical axis of the excitation light L1 reflected by the total reflection mirror 111 with the optical axis of the excitation light L2 emitted from the excitation light source 102.
  • the excitation light L1 from the reflection mirror 111 is transmitted, and the excitation light L2 from the excitation light source 102 is reflected.
  • a dichroic mirror designed to transmit light having a wavelength of 637 nm and reflect light having a wavelength of 488 nm may be used.
  • the dichroic mirror 113 is an optical element for aligning or paralleling the optical axes of the excitation lights L1 and L2 from the dichroic mirror 112 with the optical axes of the excitation light L3 emitted from the excitation light source 103.
  • the excitation light L1 from the reflection mirror 111 is transmitted, and the excitation light L3 from the excitation light source 103 is reflected.
  • a dichroic mirror designed to transmit light having a wavelength of 637 nm and light having a wavelength of 488 nm and to reflect light having a wavelength of 405 nm may be used.
  • the excitation lights L1 to L3 finally collected as light traveling in the same direction by the dichroic mirror 113 are incident on the dichroic mirror 115 through the holes 114a provided in the perforated mirror 114.
  • FIG. 2 is a diagram showing an example of a reflecting surface of the perforated mirror according to the present embodiment
  • FIG. 3 is a cross section showing dimensions when the perforated mirror according to the present embodiment is installed on an optical path of excitation light. It is a figure.
  • the perforated mirror 114 has, for example, a structure in which a hole 114a is provided substantially in the center of a circular reflecting surface.
  • the reflective surface of the perforated mirror 114 is designed to reflect, for example, at least light having a wavelength of 488 nm, which corresponds to excitation light L2.
  • the perforated mirror 114 directs at least a part of the backscattered light L12 from the spot 123a set in the microchip 120, which will be described later, in a direction different from the optical axis of the excitation lights L1 to L3.
  • the excitation lights L1 to L3 are arranged at a predetermined angle (for example, 45 degrees) with respect to the optical axis.
  • a backscattered light detection system 130 which will be described later, is arranged in the traveling direction of the backscattered light L12 reflected by the perforated mirror 114.
  • the perforated mirror 114 is arranged on the optical path of the excitation lights L1 to L3 so that the optical axis of the excitation lights L1 to L3 passes substantially in the center of the holes 114a.
  • the diameter of the hole 114a is, for example, the shortest diameter D of the hole 114a seen from the optical axis direction when the perforated mirror 114 is installed at an angle ⁇ with respect to the optical axes of the excitation lights L1 to L3.
  • the diameter may be at least larger than the diameter d of the beam cross section of the collected excitation lights L1 to L3.
  • the diameter of the beam cross section may be, for example, the diameter of a region where the beam intensity in the beam cross section is equal to or higher than a predetermined value when the beam cross section is circular.
  • the numerical aperture of the hole 114a viewed from the direction inclined by the angle ⁇ may be 0.15 or more.
  • the hole 114a is made too large, the backscattered light L12 incident on the backscattered light detection system 130 is reduced, so that the numerical aperture of the hole 114a is preferably as small as possible.
  • the shape of the reflecting surface of the perforated mirror 114 and the shape of the hole 114a are not limited to a circle, but may be an ellipse or a polygon. Further, the shape of the reflecting surface of the perforated mirror 114 and the shape of the hole 114a do not have to be similar to each other, and may be independent of each other.
  • the dichroic mirror 115 on which the excitation lights L1 to L3 that have passed through the holes 114a are incident has, for example, light having a wavelength of 637 nm corresponding to the excitation light L1 and light having a wavelength of 488 nm corresponding to the excitation light L2. It is designed to reflect light having a wavelength of 405 nm, which corresponds to the excitation light L3, and to transmit light having another wavelength. Therefore, the excitation lights L1 to L3 incident on the dichroic mirror 115 are reflected by the dichroic mirror 115 and incident on the objective lens 116.
  • a beam shaping unit for converting the excitation lights L1 to L3 into parallel light may be provided on the optical path from the excitation light sources 101 to 103 to the objective lens 116.
  • the beam shaping unit may be composed of, for example, one or more lenses, mirrors, and the like.
  • the objective lens 116 collects the incident excitation lights L1 to L3 on a predetermined spot 123a on the flow path in the microchip 120, which will be described later.
  • the spots 123a are irradiated with excitation lights L1 to L3, which are pulsed lights, while the fine particles are passing through the spot 123a, fluorescence is emitted from the fine particles, and the excitation lights L1 to L3 are the fine particles. It is scattered and scattered light is generated.
  • the component within a predetermined angle range in which the excitation light L1 to L3 travels forward in the traveling direction is referred to as forward scattered light
  • the traveling of the excitation light L1 to L3 is referred to as backscattered light L12
  • a component in a direction deviating from the optical axis of the excitation lights L1 to L3 by a predetermined angle is referred to as side scattered light.
  • the objective lens 116 has a numerical aperture corresponding to, for example, about 40 ° to 60 ° with respect to the optical axis (for example, corresponding to the predetermined angle described above).
  • the components hereinafter referred to as fluorescence L14
  • fluorescence L14 within a predetermined angle range in which the excitation lights L1 to L3 travel backward in the traveling direction and the backscattered light L12 pass through the objective lens 116 and are a dichroic mirror. It is incident on 115.
  • the fluorescence L14 passes through the dichroic mirror 115 and is incident on the fluorescence detection system 140.
  • the backscattered light L12 is reflected by the dichroic mirror 115, further reflected by the perforated mirror 114, and incident on the backscattered light detection system 130.
  • the numerical aperture of the hole 114a of the perforated mirror 114 is set to a numerical aperture of about 20 ° with respect to the optical axis (for example, NA ⁇ 0.2), and the numerical aperture of the objective lens 116 is 40 ° with respect to the optical axis.
  • the numerical aperture is about the same, the rear scattered light L12 within an angle range of about 20 ° to 40 ° with respect to the optical axis is incident on the rear scattered light detection system 130. That is, the backscattered light L12 having a donut-shaped beam profile is incident on the backscattered light detection system 130.
  • the rear-scattered light detection system 130 includes, for example, a plurality of lenses 131, 133 and 135 that shape the beam cross section of the rear-scattered light L12 reflected by the perforated mirror 114, an aperture 132 that adjusts the amount of light of the rear-scattered light L12, and the like. Detects a mask 134 that selectively transmits light of a specific wavelength among the rear-scattered light L12 (for example, light having a wavelength of 488 nm corresponding to excitation light L2), and light that is transmitted through the mask 134 and the lens 135 and incident. It is provided with an optical detector 136.
  • the diaphragm 132 may have, for example, a configuration in which a pinhole-shaped hole is provided in a light-shielding plate. This hole may be larger than the width of the hole (region where the laser intensity is reduced) in the central portion of the backscattered light L12 having a donut-shaped beam profile.
  • the photodetector 136 is composed of, for example, a two-dimensional image sensor, a photodiode, or the like, and detects the amount and size of light incident through the mask 134 and the lens 135.
  • the signal detected by the photodetector 136 is input to, for example, the analysis system 212 described later.
  • the analysis system 212 for example, the size of fine particles may be analyzed based on the input signal.
  • the fluorescence detection system 140 is, for example, a spectroscopic optical system 141 that disperses incident fluorescence L14 into dispersed light L15 for each wavelength, and a photodetector that detects the amount of dispersed light L15 for each predetermined wavelength band (also referred to as a channel). It is equipped with 142. Further, the fluorescence detection system 140 includes an imaging lens 143 that concentrates the fluorescence L14 of the collimated light transmitted through the dichroic mirror 115, and a preparative fiber 144 that guides the condensed fluorescence L14 to a predetermined position.
  • FIG. 5 shows a more detailed configuration example of the optical system from the spot 123a in the microchip 120 in FIG. 1 to the spectroscopic optical system 141.
  • the dichroic mirror 115 in FIG. 1 is omitted.
  • the fluorescence L14 radiated from the spot 123a is converted into collimated light by the objective lens 116 and then condensed by the imaging lens 143 to be focused on one end (incident end) of the preparative fiber 144. Introduced in. After that, the fluorescence L14 is guided to the spectroscopic optical system 141 by emitting light from the other end (emission end) of the preparative fiber 144.
  • FIG. 6 is a diagram showing an example of the beam diameter of the fluorescent L14 in service at the incident end of the preparative fiber 144 and the core diameter of the preparative fiber 144.
  • the aperture (core diameter) of the preparative fiber 144 also has a field aperture function of cutting stray light such as excitation light reflected by the end face of the microchip 120. Therefore, it is desirable that the core diameter of the preparative fiber 144 is as small as possible. For example, it is desirable that the core diameter of the preparative fiber 144 is a size corresponding to the flow path width of the microchip 120.
  • FIG. 7 shows an example of the spectroscopic optical system 141 according to the present embodiment.
  • the spectroscopic optical system 141 includes, for example, one or more optical elements 141a such as a prism and a diffraction grating, and emits incident fluorescent L14 at different angles for each wavelength.
  • the spectrum is dispersed on the dispersed light L15.
  • the photodetector 142 may be composed of, for example, a plurality of light receiving units that receive light for each channel.
  • the plurality of light receiving units may be arranged in one row or two or more rows in the spectral direction H1 by the spectroscopic optical system 141.
  • a photoelectric conversion element such as a photomultiplier tube can be used for each light receiving unit.
  • the photodetector 142 it is also possible to use a two-dimensional image sensor or the like instead of a plurality of light receiving units such as a photomultiplier tube array.
  • a signal indicating the amount of light of the fluorescence L14 for each channel detected by the photodetector 142 is input to, for example, an analysis system 212 described later.
  • the analysis system 212 for example, component analysis and morphological analysis of fine particles may be executed based on the input signal.
  • FIG. 8 is a block diagram showing a schematic configuration example of the information processing system according to the present embodiment.
  • the information processing system includes, for example, an analysis system 212 that acquires a signal from the photodetector 142 and / or the photodetector 136 and analyzes fine particles based on the acquired signal.
  • the signals generated by the photodetectors 136 and 142 may be various signals such as image data and optical signal information.
  • the analysis system 212 may be a local PC (personal computer), a cloud server, a part of the local PC, and a part of the cloud server.
  • the cell analyzer 1 when the cell analyzer 1 is a sorter, the cell analyzer 1 may include a sorting control unit that controls the sorting of fine particles (for example, cells) based on the analysis result.
  • the forward scattered light may be used to specify the timing at which the fine particles pass through the spot 123a set on the flow path in the microchip 120. Therefore, in the present embodiment, the forward scattered light detection system 160 is provided.
  • the light L16 that travels forward in the traveling direction of the excitation lights L1 to L3 from the fine particles includes forward scattered light and components within a predetermined angle range that travels forward in the traveling direction of the excitation lights L1 to L3 among the fluorescence emitted from the fine particles. And are included.
  • the filter 151 arranged on the optical path of the excitation lights L1 to L3 on the downstream side of the microchip 120 includes, for example, light having a wavelength of 637 nm corresponding to the excitation light L1 (forward scattered light L17).
  • light having a wavelength of 488 nm (forward scattered light L18) corresponding to the excitation light L2 is selectively transmitted, and light having other wavelengths is blocked.
  • FIG. 9 is a schematic diagram showing a filter installed with respect to the optical axis of the light traveling forward in the traveling direction of the excitation light from the fine particles.
  • the filter 151 is arranged so as to be inclined with respect to the optical axis of the light L16. This prevents the return light of the light L16 reflected by the filter 151 from entering the backscattered light detection system 130 or the like via the objective lens 116 or the like.
  • the forward scattered light L17 and L18 that have passed through the filter 151 are converted into parallel light by passing through the collimating lens 152, then reflected in a predetermined direction by the total reflection mirror 153 and incident on the forward scattered light detection system 160. ..
  • the forward scatter light detection system 160 includes a lens 161, a dichroic mirror 162a, a total reflection mirror 162b, apertures 163a and 163b, lenses 164a and 164b, filters 165a and 165b, diffraction gratings 166a and 166b, and photodetection. It is provided with vessels 167a and 167b.
  • the dichroic mirror 162a reflects the forward scattered light L17, which is the scattered light of the excitation light L1, among the forward scattered lights L17 and L18 reflected by the fully reflective mirror 153, and produces the forward scattered light L18, which is the scattered light of the excitation light L2. It is designed to be transparent.
  • the lens 161 and the lens 164a function as an optical system for shaping the beam cross section of the forward scattered light L17 traveling on the optical path sandwiched between them.
  • the diaphragm 163a adjusts the amount of light of the forward scattered light L17 incident on the photodetector 167a.
  • the filter 165a and the diffraction grating 166a function as an optical filter that enhances the purity of the forward scattered light L17 in the light incident on the photodetector 167a.
  • the photodetector 167a is composed of, for example, a photodiode, and detects the incident of the forward scattered light L17.
  • the lens 161 and the lens 164b function as an optical system for shaping the beam cross section of the forward scattered light L18 traveling on the optical path sandwiched between them.
  • the diaphragm 163b adjusts the amount of light of the forward scattered light L18 incident on the photodetector 167b.
  • the filter 165b and the diffraction grating 166b function as an optical filter that enhances the purity of the forward scattered light L18 in the light incident on the photodetector 167b.
  • the photodetector 167b is composed of, for example, a photodiode, and detects the incident of the forward scattered light L18.
  • a detection system for detecting the forward scattered light L17 (lens 161 and 164a, aperture 163a, filter 165a, diffraction grating 166a, and light detector 167a). And a detection system (lens 161 and 164b, aperture 163b, filter 165b, diffraction grating 166b, and light detector 167b) for detecting forward scattered light L18.
  • the timing detected by one of the detection systems for example, the detection system for detecting the forward scattered light L18
  • the other detection system for example, the detection system for detecting the forward scattered light L17 It is possible to compensate at the timing.
  • the configuration is not limited to this, and for example, one of the detection systems may be omitted.
  • the timing referred to here may be the timing at which the fine particles pass through the spot 123a set on the flow path in the microchip 120.
  • an optical system for irradiating the spot 123a with the excitation light L1 to L3 and a detection system for detecting the fluorescence L14 and the backward scattered light L12 from the spot 123a, that is, an excitation light source may be mounted on the same base 100.
  • a detection system for detecting the forward scattered light L17 and L18 from the spot 123a that is, a backscattered light detection system 130, a fluorescence detection system 140, a filter 151, a total reflection mirror 153, and a forward scattered light detection.
  • the system 160 may be mounted on the same base 150 different from the base 100. Further, the base 100 and the base 150 may be aligned with each other.
  • FIG. 10 is a diagram schematically showing a schematic configuration of a microchip according to the present embodiment.
  • a sample liquid introduction flow path 121 into which the sample liquid 126 containing fine particles is introduced and a pair of sheath liquid introduction flows into which the sheath liquid 127 is introduced.
  • Roads 122a and 122b are provided.
  • the fine particles may include, for example, cells, cell groups, tissues, and the like when the object to be observed is a biological substance.
  • the observation target is not limited to these, and various fine particles can be observed.
  • the sheath liquid introduction flow paths 122a and 122b merge with the sample liquid introduction flow path 121 from both sides, and one merging flow path 123 is provided on the downstream side of the merging point. Then, in the merging flow path 123, the sample liquid 126 is surrounded by the sheath liquid 127 so that the liquid flows in a state where a laminar flow is formed. As a result, the fine particles in the sample liquid 126 are allowed to flow in substantially one row in the flow direction.
  • 125b and 125b are provided, and all of them communicate with the merging flow path 123.
  • the downstream ends of the waste flow paths 125a and 125b are connected to, for example, a waste liquid tank.
  • this microchip 120 individual fine particles are detected in the merging flow path 123, and as a result, only the fine particles determined to be collected are drawn into the negative pressure suction unit 124, and the other fine particles are removed. It is discharged from the disposal channels 125a and 125b.
  • the structure of the negative pressure suction unit 124 is not particularly limited as long as it can suck the fine particles to be collected at a predetermined timing.
  • the negative pressure suction unit 124 communicates with the confluence flow path 123. It can be composed of a suction flow path 124a to be formed, a pressure chamber 124b formed in a part of the suction flow path 124a, and an actuator 124c whose volume in the pressure chamber 124b can be expanded at an arbitrary timing. It is desirable that the downstream end of the suction flow path 124a can be opened and closed by a valve (not shown) or the like.
  • the pressure chamber 124b is connected to an actuator 124c such as a piezo element via a diaphragm.
  • the material for forming the microchip 120 examples include polycarbonate, cycloolefin polymer, polypropylene, PDMS (polydimethylsiloxane), glass, and silicon.
  • it is preferably formed of a polymer material such as polycarbonate, cycloolefin polymer, or polypropylene because it has excellent processability and can be replicated at low cost using a molding apparatus. In this way, the microchip 120 can be manufactured at low cost by adopting the structure in which the plastic molded substrates are bonded together.
  • the method of supplying fine particles to the spot on the flow path in the present embodiment is not limited to the microchip method, and various methods such as a droplet method, a cuvette method, and a flow cell method are adopted. It is possible to do.
  • FIG. 11 is a diagram for explaining a case where the fluorescence and the backscattered light according to the comparative example are not separated
  • FIG. 12 is a diagram for explaining a case where the fluorescence and the backscattered light according to the present embodiment are separated. It is a figure of.
  • the fluorescence L14 and the backscattered light L12 are not separated but are reflected by the perforated mirror 114 and are incident on the detection system.
  • the detection system for detecting the backscattered light L12 and the detection system for detecting the fluorescence L14 are arranged on the optical axis of the fluorescence L14 and the backscattered light L12 reflected by the perforated mirror 114. And.
  • the fluorescence L14 and the backscattered light L12 are not separated in this way, the fluorescence L14 near the optical axis having a relatively high beam intensity will pass through the hole 114a of the perforated mirror 114. Therefore, the sensitivity of the detection system to the fluorescence L14 is lowered, and the detection efficiency and the detection accuracy are lowered.
  • the fluorescence L14 and the backscattered light L12 that have passed through the hole 114a may be absorbed by, for example, a beam damper (not shown).
  • the optical axis having a relatively high beam intensity is configured to be separated by the dichroic mirror 115 before being reflected by the fluorescence L14, the backscattered light L12, and the perforated mirror 114. It is possible to make the nearby fluorescence L14 incident on the fluorescence detection system 140 without discarding it. As a result, it is possible to suppress a decrease in the sensitivity of the fluorescence detection system 140 with respect to the fluorescence L14, so that it is possible to suppress a decrease in detection efficiency and a decrease in detection accuracy.
  • the excitation lights L1 to L3 are irradiated substantially perpendicular to the incident surface of the microchip 120. With such a configuration, it is difficult to observe the laterally scattered light among the scattered light scattered by the fine particles. Therefore, in the present embodiment, the forward scattered light L17 and L18 and the backscattered light L12 are observed, and among these, the backscattered light L12 is observed as the return light via the objective lens 116.
  • the objective lens 116 is required to have optical stability that can maintain its optical characteristics even when it is irradiated with excitation lights L1 to L3 having a strong laser intensity. For example, even if strong ultraviolet light having a wavelength of 450 nm or less (for example, excitation light L3) is irradiated, high optical stability is required so that the optical characteristics are maintained to the extent that the observation of backscattered light L12 is not hindered. Be done.
  • a bonded lens formed by adhering a plurality of lenses can be used in order to enable aberration correction.
  • adhesives are usually used for bonded lenses to fix individual lenses. Therefore, when the light source of the flow cytometer contains light rays in the ultraviolet (wavelength 450 nm or less) region (for example, excitation light L3), the adhesive at the lens junction is burnt or emitted from the adhesive to the lens surface. There is a possibility that the attached outgas may be burnt and the optical characteristics of the bonded lens may be deteriorated.
  • the objective lens 116 having a novel structure of introducing a junction division group and a telephoto configuration is used.
  • the objective lens 116 having such a structure it is possible to achieve the effect of correcting chromatic aberration and avoiding burning of the adhesive, etc., and at the same time, the effect of reducing the cost by reducing the number of mechanical parts and the number of lenses. It will be possible.
  • FIG. 13 is a cross-sectional view showing a schematic configuration example of the objective lens according to the present embodiment. Note that FIG. 13 shows a cross-sectional structure when the objective lens 116 is cut on a surface including the optical axes of the excitation lights L1 to L3. Further, FIG. 14 is an optical path diagram showing a light beam of the objective lens shown in FIG.
  • the objective lens 116 is an infinite correction objective lens.
  • the objective lens 116 is a first lens having positive power (hereinafter, hereinafter, in order from the incident side of the excitation lights L1 to L3 and the emission side (infinity side) of the fluorescent L14 and the backward scattered light L12). It consists of a first positive lens (21), a positive power second lens (hereinafter referred to as the second positive lens) 22 and a negative power third lens (hereinafter referred to as the third negative lens) 23, and is a positive power junction as a whole.
  • the split group 24, the positive power fourth lens (hereinafter referred to as the fourth positive lens) 25, and the negative power fifth lens (hereinafter referred to as the fifth negative lens) 26 are composed of the positive power junction split group 27 as a whole. It is composed of a positive power sixth lens (hereinafter referred to as a sixth positive lens) 28.
  • the objective lens 116 has, for example, a focal length of 10 mm, a numerical aperture of NA 0.75, and an objective field of view of ⁇ 0.5 mm, and covers the wavelength bands of excitation light L1 to L3 and fluorescence L13 of 405 to 850 nm. Since the objective lens 116 having such a configuration has a narrow field of view, the priority of correcting aberrations (magnification color, curvature of field, distortion) depending on the angle of view is not high. However, since the numerical aperture NA is large, it is necessary to sufficiently correct the aberration (spherical surface / coma) depending on the aperture 10 shown in FIG. Further, since the wavelength band is wide, it is necessary to sufficiently correct the axial chromatic aberration.
  • the coupling efficiency of the semaphore 144 is defined by (the amount of signals entering the core of the semaphore 144) / (the total amount of signals on the incident end face of the semaphore 144), while as described above. It is desirable that the core diameter of the semaphore fiber 144 is as small as possible.
  • the amount of signal entering the core (the amount of light of the fluorescence L14) is reduced, and the coupling efficiency is lowered. .. In that case, there arises a problem that the detection sensitivity of the cell analyzer 1 is lowered.
  • the excitation lights L1 to L3 contain ultraviolet rays (wavelength 450 nm or less), the ultraviolet curable adhesive used for the joint surface is burnt. As a result, the transmittance may decrease with continuous use, and the detection sensitivity of the cell analyzer 1 may decrease.
  • the junction division group 24 including the second positive lens 22 and the third negative lens 23 and the junction division group 27 including the fourth positive lens 25 and the fifth negative lens 26 are used.
  • it also avoids burning of the bonding adhesive due to the use of an ultraviolet excitation laser (for example, corresponding to the excitation light source 103).
  • junction division groups 24 and 27 By using two junction division groups (junction division groups 24 and 27) as in the present embodiment, axial chromatic aberration is satisfactorily corrected while suppressing an increase in size and cost of the optical system. It becomes possible.
  • this description does not exclude from the technical scope of the present disclosure that there are one or three or more junction division groups, and the number of junction division groups may be one or three. It may be one or more.
  • the general junction group has three surfaces that contribute to aberration correction, whereas the junction division group has four surfaces. Therefore, the degree of freedom due to the four contributing surfaces can be allocated to the above-mentioned spherical / coma aberration correction. As a result, it is possible to realize good aberration correction with a small number of components of 6 elements in 6 groups, and thus it is possible to reduce the cost.
  • the objective lens 116 is previously made into a focused ray by the weak refractive power of the first positive lens 21 having a positive power, and then guides the ray to the junction division groups 24 and 27. There is. As a result, the positive power of each of the junction division groups 24 and 27 can be weakened, so that the occurrence of aberration can be suppressed in the entire optical system.
  • the objective lens 116 has a structure close to that of the telephoto type. As a result, it is possible to gradually reduce the outer shape of the lens from the emission side (infinity side) of the fluorescence L14 and the backscattered light L12 on the incident side of the excitation lights L1 to L3 toward the object side.
  • the lens barrel can be designed so that the lens is fitted into one lens frame 10. As a result, it is possible to reduce the cost of mechanical parts.
  • the relative positions of the joint division surfaces in the joint division groups 24 and 27 may be determined by marginal contacts in which the curved surfaces of the polished surfaces are in direct contact with each other. The reason is as follows.
  • the overall performance is exhibited by canceling the aberrations between the joint dividing surfaces. Therefore, if the joint division surfaces have eccentricity during manufacturing, the performance is large. May decrease. In this respect, by directly contacting the curved surfaces of the polished surfaces with each other, the relative eccentricity between the surfaces can be set to zero even if an error in the external dimension of the lens occurs.
  • the positive lens (second lens 22 and / or fourth lens 25) constituting at least one of the two junction division groups (24, 27) according to the present embodiment has a refractive index Nd of 1. It may be 6 or less, the Abbe number ⁇ d is 65 or more, and the partial dispersion ratio ⁇ gF may be 0.55 or less.
  • the refractive index Nd in this description is the refractive index at the d-line 587.56 nm
  • the Abbe number ⁇ d is the Abbe number at the d-line 587.56 nm
  • the partial dispersion ratio ⁇ gF is the g-line 435.834 nm. It is a partial dispersion ratio defined by the F-line 486.133 nm.
  • FIG. 15 is a cross-sectional view showing a schematic configuration example of the objective lens according to the modified example
  • FIG. 16 is an optical path diagram showing a light ray of the objective lens shown in FIG.
  • the objective lens 416 is a negative power first lens (hereinafter referred to as a first negative lens) 41 and a positive power second lens (hereinafter referred to as a second positive lens). ) 42 as a whole, consisting of a negative power junction division group 43 and positive power third to seventh lenses (hereinafter referred to as third to seventh positive lenses) 44 to 48.
  • a negative power first lens hereinafter referred to as a first negative lens
  • a positive power second lens hereinafter referred to as a second positive lens
  • ) 42 as a whole, consisting of a negative power junction division group 43 and positive power third to seventh lenses (hereinafter referred to as third to seventh positive lenses) 44 to 48.
  • the curvature of field is corrected by using the first negative lens 41 having a low light beam height as a negative lens with strong power. Has been done. Then, since the light rays inevitably diverge from the first negative lens 41 to the third positive lens 44, the outer shape of the lens increases toward the middle and then decreases from the fourth positive lens 45 to the seventh positive lens 48. Therefore, two parts are required, a first lens frame 50 that holds the first negative lens 41 to the third positive lens 44, and a second lens frame 60 that holds the fourth positive lens 45 to the seventh positive lens 48. The number of parts increases and the assembly process becomes complicated, resulting in an increase in manufacturing costs.
  • the introduction of the junction division group and the telephoto configuration have the effects of correcting chromatic aberration and avoiding burning of the adhesive or the like. It is possible to achieve the effect of cost reduction by reducing the number of mechanical parts and the number of lenses.
  • the lens frame that holds the above-mentioned optical system will be described.
  • the sixth positive lens 28 arranged on the microchip 120 side is fitted into the lens frame 10 through the opening 12 on the microchip 120 side.
  • the first positive lens 21, the second positive lens 22, the third negative lens 23, the fourth positive lens 25, and the fifth negative lens 26 are fluorescent on the incident side of the excitation lights L1 to L3 in ascending order of diameter. It is fitted into the lens frame 10 through the opening 11 on the emission side (infinity side) of L14 and the rearward scattered light L12.
  • the first positive lens 21, the second positive lens 22, the third negative lens 23, the fourth positive lens 25, the fifth negative lens 26, and the sixth positive lens 28 are the excitation lights L1 to L3. They are arranged along the optical axis in descending order of diameter in the direction perpendicular to the optical axis.
  • the inside of the lens frame 10 is stepwise reduced in diameter according to the diameters of the first positive lens 21, the second positive lens 22, the third negative lens 23, the fourth positive lens 25, and the fifth negative lens 26.
  • the fifth negative lens 26 which is first fitted from the aperture 11 side, abuts on the abutting portion 13 in the lens frame 10 and makes marginal contact with the fourth positive lens 25, so that the fifth negative lens 26 is in the lens frame 10. It is fixed.
  • the fourth positive lens 25 is fixed in the lens frame 10 by making marginal contact with the fifth negative lens 26 and abutting with the spacing ring 34 that functions as a spacer.
  • the diameters of the fourth positive lens 25 and the fifth negative lens 26 are about the same, and the diameter of the portion where the fourth positive lens 25 and the fifth negative lens 26 are located inside the lens frame 10 is the fourth.
  • the positive lens 25 and the fifth negative lens 26 are designed to fit exactly.
  • the spacing ring 34 has a ring shape with an opening at the center, and after fitting the fourth positive lens 25 into the lens frame 10, before fitting the third negative lens 23 into the lens frame 10. , It is fitted in the lens frame 10.
  • the outer diameter of the spacing ring 34 may be, for example, about the same as that of the third negative lens 23 and the second positive lens 22.
  • the third negative lens 23 is fixed in the lens frame 10 by abutting on the spacing ring 34 fitted in the lens frame 10 and making marginal contact with the second positive lens 22. At that time, the spacing ring 34 is fixed in the lens frame 10 by being sandwiched between the fourth positive lens 25 and the third negative lens 23.
  • the second positive lens 22 is fixed in the lens frame 10 by making marginal contact with the third negative lens 23 and abutting with the spacing ring 32 that functions as a spacer.
  • the diameters of the second positive lens 22 and the third negative lens 23 are about the same, and the diameter of the portion where the second positive lens 22 and the third negative lens 23 are located inside the lens frame 10 is the second.
  • the positive lens 22 and the third negative lens 23 are designed to fit exactly.
  • the spacing ring 32 has a ring shape with an opening at the center, and after fitting the second positive lens 22 into the lens frame 10, before fitting the first positive lens 21 into the lens frame 10. , It is fitted in the lens frame 10.
  • the outer diameter of the spacing ring 32 may be, for example, about the same as that of the first positive lens 21 or about the same as that of the second positive lens 22.
  • the first positive lens 21 comes into contact with the spacing ring 32 fitted in the lens frame 10, and the mounting screw 30 having an opening in the center is turned into the screw frame provided on the opening 11 side to turn the first positive lens 21. By contacting with 21, it is fixed in the lens frame 10.
  • a metal or alloy such as aluminum or brass can be used.
  • metals such as aluminum and copper, alloys and the like can be used for the spacing rings 32 and 34 and the mounting screws 30.
  • the material is not limited to these materials, and various materials can be adopted in consideration of price, ease of processing, durability, and the like.
  • an air hole 17 for allowing internal air to escape when the fifth negative lens 26 or the sixth positive lens 28 is fitted into the lens frame 10 and a third negative lens 23 are attached to the lens frame 10. Even if an air hole 16 for letting out the internal air when fitting into the lens and an air hole 15 for letting out the internal air when fitting the first positive lens 21 into the lens frame 10 are provided. Good.
  • a plurality of lenses (first positive lens 21, second positive lens 22, third negative lens 23, fourth positive lens 25, and fifth negative lens 26).
  • the entire lens is sandwiched between the lens frame 10 and the mounting screw 30, and each lens is fixed by the marginal contact between the lenses and the contact between the spacing ring 32/34. It is possible to fix the relative position between the faces.
  • the second positive lens 22 and the third negative lens 23 and the fourth positive lens 25 and the fifth negative lens 26 are positioned with each other by marginal contacts that come into contact with each other. Further, the first positive lens 21 and the second positive lens 22 and the third negative lens 23 and the fourth positive lens 25 are positioned with each other by abutting with the spacing rings 32 and 34 interposed therein. Further, in the first positive lens 21, the second positive lens 22, the third negative lens 23, the fourth positive lens 25, and the fifth negative lens 26 as a whole, the fifth negative lens 26 hits the contact portion 14 of the lens frame 10.
  • the first positive lens 21 is fixed in the lens frame 10 by being in contact with the mounting screw 30 and being urged by the mounting screw 30.
  • the sixth positive lens 28 fitted from the opening 12 side is held by the lens frame 10 by abutting on the abutting portion 14 in the lens frame 10. At that time, since the sixth positive lens 28 is not sealed by the lens frame 10, it may be fixed to the lens frame 10 with an adhesive or the like. However, the present invention is not limited to this, and the sixth positive lens 28 may be fixed to the lens frame 10 by covering the opening 12 with a cap having an opening at the center.
  • the objective lens 116 can hold a plurality of lenses (21, 22, 23, 25, 26 and 28) in one lens frame 10, the cost due to the reduction in the number of parts is reduced. It is also possible to achieve the effects of reduction and simplification of the assembly process.
  • junction division groups 24 and 27 since there are two junction division groups (junction division groups 24 and 27), axial chromatic aberration is suppressed while suppressing an increase in size and cost of the optical system. It is possible to make good corrections, and for example, it is possible to reduce the number of lens points as compared with the objective lens 416 according to the modified example, so that the cost can be reduced by reducing the number of parts and the assembly process can be simplified. It is also possible to achieve the effect.
  • FIG. 17 is a cross-sectional view showing a schematic configuration example of the objective lens according to the first specific example.
  • FIG. 18 is a cross-sectional view showing a schematic configuration example of an imaging lens used in combination with the objective lens according to the first to third specific examples.
  • Table 1 shows an example of lens data of each lens constituting the objective lens 116A according to the first specific example, and Table 2 shows an example of lens data of the imaging lens 143.
  • the focal length fo of the objective lens 116A is 10 mm
  • the opening number NA of the objective lens 116A on the object side is 0.65
  • the magnification ⁇ is 6.5
  • G13 (S8) is illustrated in which the partial dispersion ratio ⁇ gF of the surface glass material) is 0.5392
  • the focal length fi of the imaging lens 143 is 65 mm
  • the distance between the objective lens 116A and the imaging lens 143 is 66.0 mm.
  • S indicates the plane number
  • R indicates the radius of curvature
  • Nd indicates the refractive index with respect to the d line
  • ⁇ d indicates the Abbe number with respect to the d line.
  • S1 surface the surface of the surface number S1 (hereinafter referred to as the S1 surface; the same applies to the other surface numbers) is the object surface of the fine particles to be observed
  • the S1 to S3 surfaces are on the microchip 120 side.
  • the surface is the surface
  • the S4 surface is the incident surface of the objective lens 116A
  • the S12 surface is the exit surface of the objective lens 116A.
  • the S1 surface is the entrance surface of the imaging lens 143
  • the S3 surface is the exit surface of the imaging lens 143.
  • the objective lens 116A is a positive lens G11 having a positive refractive power in order from the upstream side, that is, the side closer to the microchip 120. It is composed of a negative lens G12 having a negative refractive power, a positive lens G13 having a positive refractive power, and a positive lens G14 having a positive refractive power.
  • the negative lens G12 and the positive lens G13 form a junction division group GR11.
  • the positive lens G11 is a biconvex lens
  • the negative lens G12 is a biconcave lens
  • the positive lens G13 is a biconvex lens
  • the positive lens G14 is a biconvex lens.
  • the imaging lens 143 is used integrally with the objective lens 116A.
  • the imaging lens 43 is composed of, for example, a junction lens of a positive lens G1 having a positive refractive power and a negative lens G2 having a negative refractive power.
  • the positive lens G1 is, for example, a biconvex lens having a partial dispersion ratio ⁇ gF of 0.5375
  • the negative lens G2 is, for example, a meniscus lens having a concave surface facing the object side.
  • FIGS. 19 to 21 are views showing an example of longitudinal aberration of an optical system in which an objective lens and an imaging lens according to a first specific example are combined
  • FIGS. 22 to 25 are objectives according to the first specific example. It is a figure which shows an example of the lateral aberration of the optical system which combined the lens and the imaging lens.
  • the aberration is satisfactorily corrected in a wide wavelength band from 404.656 nm to 852.110 nm. Is possible.
  • FIG. 26 is a cross-sectional view showing a schematic configuration example of the objective lens according to the second specific example.
  • the imaging lens 143 may be the same as the imaging lens 143 illustrated with reference to FIGS. 18 and 2 above. Further, Table 3 below shows an example of lens data of each lens constituting the objective lens 116B according to the second specific example.
  • the focal length fo of the objective lens 116B is 10 mm
  • the numerical aperture NA of the objective lens 116B on the object side is 0.75
  • the magnification ⁇ is 6.5
  • the portion of G23 glass material on the S8 surface.
  • An example is illustrated in which the dispersion ratio ⁇ gF and the partial dispersion ratio ⁇ gF of G25 (glass material on the S12 surface) are both 0.5375, and the distance between the objective lens 116B and the imaging lens 143 is 66.0 mm.
  • the S1 surface is the object surface of the fine particles to be observed
  • the S1 to S3 surfaces are the surfaces on the microchip 120 side
  • the S4 surface is the incident surface of the objective lens 116B
  • the S16 surface Is the exit surface of the objective lens 116B.
  • the objective lens 116B has the positive lens G21 having a positive refractive power and the negative refractive power in this order from the upstream side, that is, the side closer to the microchip 120. It is composed of a negative lens G22 having a positive refractive power, a positive lens G23 having a positive refractive power, a negative lens G24 having a negative refractive power, a positive lens G25 having a positive refractive power, and a positive lens G26 having a positive refractive power. There is.
  • the negative lens G22 and the positive lens G23 form a junction division group GR21
  • the negative lens G24 and the positive lens G25 form a junction division group GR22.
  • the positive lens G21 is a meniscus lens with a concave surface facing the microchip 120 side
  • the negative lens G22 is a meniscus lens with a concave surface facing the preparative fiber 144 side.
  • the positive lens G23 is a biconvex lens
  • the negative lens G24 is a biconcave lens
  • the positive lens G25 is a biconvex lens
  • the positive lens G26 is a meniscus lens with a concave surface facing the microchip 120 side.
  • FIGS. 27 to 29 are views showing an example of longitudinal aberration of an optical system in which an objective lens and an imaging lens according to a second specific example are combined
  • FIGS. 30 to 33 are objectives according to the second specific example. It is a figure which shows an example of the lateral aberration of the optical system which combined the lens and the imaging lens.
  • the objective lens 116B according to the second specific example can satisfactorily correct the aberration in a wide wavelength band from 404.656 nm to 852.110 nm. It is possible.
  • FIG. 34 is a cross-sectional view showing a schematic configuration example of the objective lens according to the third specific example.
  • the imaging lens 143 may be the same as the imaging lens 143 illustrated with reference to FIGS. 18 and 2 above. Further, Table 4 below shows an example of lens data of each lens constituting the objective lens 416A according to the third specific example.
  • the focal length fo of the objective lens 416A is 10 mm
  • the numerical aperture NA of the objective lens 416A on the object side is 0.85
  • the magnification ⁇ is 6.5
  • the portion of G33 glass material on the S8 surface.
  • the dispersion ratio ⁇ gF and the partial dispersion ratio ⁇ gF of G35 are both 0.5340
  • the distance between the objective lens 416A and the imaging lens 143 is 66.0 mm.
  • the S1 surface is the object surface of the fine particles to be observed
  • the S1 to S3 surfaces are the surfaces on the microchip 120 side
  • the S4 surface is the incident surface of the objective lens 416A
  • the S20 surface Is the exit surface of the objective lens 416A.
  • the objective lens 416A has a positive lens G31 having a positive refractive force and a negative refractive force in this order from the upstream side, that is, the side closer to the microchip 120. It is composed of a negative lens G32 having a positive refractive power, a positive lens G33 having a positive refractive power, a negative lens G34 having a negative refractive power, a positive lens G35 having a positive refractive power, and a positive lens G36 having a positive refractive power.
  • the negative lens G32 and the positive lens G33 form a junction division group GR31
  • the negative lens G34 and the positive lens G35 form a junction division group GR32
  • the positive lens G37 and the negative lens G38 form a junction division group GR33.
  • the positive lens G31 is, for example, a meniscus lens having a concave surface facing the microchip 120 side.
  • the negative lens G32 is, for example, a meniscus lens having a concave surface facing the preparative fiber 144 side
  • the positive lens G33 is, for example, a biconvex lens.
  • the negative lens G34 is, for example, a meniscus lens having a concave surface facing the preparative fiber 144 side
  • the positive lens G35 is, for example, a biconvex lens.
  • the positive lens G36 is, for example, a biconvex lens.
  • the positive lens G37 is, for example, a biconvex lens
  • the negative lens G38 is, for example, a biconcave lens
  • the amount of light of the fluorescence L14 and the backscattered light L12 can be increased by increasing the numerical aperture NA, so that the signal-to-noise ratio can be improved.
  • the junction division group GR33 is used as the second lens group 225 including the positive lens G37 and the negative lens G38.
  • the positive spherical aberration generated by the negative refractive power of the negative lens G38 can cancel the negative spherical aberration generated by the positive refractive power of the positive lenses G31, G33, G35, G36 and G37. Therefore, it is possible to reduce the spherical aberration as a whole.
  • the aplanatic property is strengthened in order to suppress the aberration.
  • the principal point on the object side moves to the image side (in this example, the preparative fiber 144 side), resulting in a so-called telephoto refractive power arrangement.
  • the working distance becomes short, it is necessary to shorten the distance between the objective lens 116/416 and the microchip 120.
  • the lens barrel of the objective lens 116/416 and the mechanical components around the microchip 120 may interfere with each other.
  • the junction division group GR33 including the positive lens G37 and the negative lens G38 is provided on the preparative fiber 144 side.
  • the negative refractive power of the negative lens G38 makes it possible to relax the telephoto configuration and bring it closer to the retrofocus configuration, and to secure a working distance. Therefore, the lens barrel and microchip 120 of the objective lens 116/416 Interference with peripheral mechanical parts can be suppressed.
  • the Petzval coefficient increases positively and negative curvature of field occurs. ..
  • a method for correcting this a method of using a glass material having a high refractive index for a lens having a positive refractive power and a glass material having a low refractive index for a lens having a negative refractive power can be considered.
  • the refractive index of generally available glass materials is about 1.40 to 2.15, and it is difficult to give a sufficient difference in the refractive index.
  • a glass material having a low refractive index and a large Abbe number ⁇ d that is, a small dispersion
  • a lens having a positive refractive power there is a need. Therefore, it is difficult to sufficiently suppress negative curvature of field by the method using glass materials having different refractive indexes.
  • a junction division group GR33 composed of a positive lens G37 and a negative lens G38 is provided.
  • the negative Petzval coefficient generated by the strong negative refractive power of the negative lens G38 can cancel the positive Petzval coefficient generated by the positive refractive power of the positive lenses G31, G33, G35, G36 and G37. Therefore, it is possible to sufficiently suppress the negative curvature of field.
  • FIGS. 35 to 37 are diagrams showing an example of the longitudinal aberration of the optical system in which the objective lens and the imaging lens according to the third specific example are combined
  • FIGS. 38 to 41 are diagrams showing an example of the longitudinal aberration of the optical system according to the third specific example. It is a figure which shows an example of the lateral aberration of the optical system which combined the lens and the imaging lens.
  • the objective lens 416A according to the third specific example can satisfactorily correct the aberration in a wide wavelength band from 404.656 nm to 852.110 nm. It is possible.
  • the present technology can also have the following configurations.
  • An excitation light source that emits excitation light with a wavelength of at least 450 nanometers or less
  • a lens structure that collects the excitation light at a predetermined position
  • a fluorescence detection system that detects fluorescence emitted from the particles when the particles existing at the predetermined positions are excited by the excitation light.
  • a scattered light detection system that detects scattered light generated by scattering the excitation light by the particles existing at the predetermined positions, and a scattered light detection system.
  • the lens structure includes a plurality of lenses arranged along the optical axis of the excitation light, and a lens frame for holding the plurality of lenses.
  • An optical measuring device in which a position in the lens frame is determined by abutting at least one of the plurality of lenses on a lens adjacent to the lens.
  • the lens structure further comprises at least one spacing ring interposed between the plurality of lenses.
  • the position in the lens frame is determined by abutting at least one of the plurality of lenses with the spacing ring interposed between the lens and the lens adjacent to the lens (1) or.
  • the plurality of lenses include at least one junction division group including a positive lens having a positive refractive power and a negative lens having a negative refractive power.
  • the optical measuring apparatus according to any one of (1) to (3), wherein the positive lens and the negative lens constituting the junction division group are in contact with each other. (5) The optical measuring device according to (4), wherein the at least one junction division group is one of the junction division groups. (6) The optical measuring device according to (4), wherein the at least one junction division group is two of the junction division groups. (7) The optical measuring device according to (4) above, wherein the at least one junction division group is three of the junction division groups. (8) The positive lens constituting at least one of the at least one junction division group has a refractive index of 1.6 or less, an Abbe number of 65 or more, and a partial dispersion ratio of 0.55 or less. The optical measuring apparatus according to any one of (4) to (7).
  • the plurality of lenses The first single lens with positive refractive power, A second single lens with positive refractive power, Including The optical measuring apparatus according to any one of (4) to (8), wherein the first single lens and the second single lens are arranged at positions sandwiching at least one junction division group.
  • the at least one junction division group includes two or more of the junction division groups.
  • the optical measuring apparatus according to any one of (1) to (10), wherein the plurality of lenses are arranged along the optical axis of the excitation light in descending order of diameter in a direction perpendicular to the optical axis. .. (12) The optical measuring device according to (11) above, wherein the lens frame is a single member. (13) The optical measuring apparatus according to any one of (1) to (12) above, wherein the scattered light is backscattered light propagating from the predetermined position along the optical path of the excitation light. (14) The optical measuring device according to any one of (1) to (13) above, wherein an adhesive is not used for fixing the plurality of lenses.
  • a lens structure that collects excitation light emitted from an excitation light source that emits excitation light having a wavelength of at least 450 nanometers or less at a predetermined position.
  • a plurality of lenses arranged along the optical axis of the excitation light, and A lens frame that holds the plurality of lenses and With A lens structure in which a position in the lens frame is determined by abutting at least one of the plurality of lenses on a lens adjacent to the lens.

Abstract

La présente invention élimine la détérioration des caractéristiques optiques. Un dispositif de mesure optique selon un mode de réalisation de la présente invention comprend : des sources de lumière d'excitation (101-103) qui émettent au moins une lumière d'excitation dont la longueur d'onde ne dépasse pas 450 nanomètres ; une structure de lentille (116) qui condense la lumière d'excitation à une position prescrite ; un système de détection de fluorescence (140) qui détecte la fluorescence émise par les particules présentes à la position prescrite résultant de l'excitation des particules par la lumière d'excitation ; et un système de détection de lumière diffusée (130) qui détecte la lumière diffusée générée par la lumière d'excitation diffusée par les particules présentes à la position prescrite. La structure de lentilles comprend : une pluralité de lentilles (21, 22, 23, 25, 26, 28) disposées le long de l'axe optique de la lumière d'excitation ; et un cadre de lentilles (10) maintenant la pluralité de lentilles. Pour au moins une lentille parmi la pluralité de lentilles, la position de ladite lentille se trouvant à l'intérieur du cadre de la lentille est déterminée par la mise en butée de la lentille contre une lentille adjacente.
PCT/JP2020/040090 2019-11-06 2020-10-26 Dispositif de mesure optique et structure de lentille WO2021090720A1 (fr)

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JP2021554896A JPWO2021090720A1 (fr) 2019-11-06 2020-10-26
US17/755,387 US20220404262A1 (en) 2019-11-06 2020-10-26 Optical measurement device and lens structure

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JP2019-201745 2019-11-06
JP2020132111 2020-08-04
JP2020-132111 2020-08-04

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004219608A (ja) * 2003-01-14 2004-08-05 Kurobane Nikon:Kk 対物レンズ及びこの対物レンズを備える顕微鏡
JP4252447B2 (ja) * 2001-06-22 2009-04-08 カール ツァイス イェナ ゲーエムベーハー 対物レンズ
JP2013152484A (ja) * 2007-07-17 2013-08-08 Olympus Corp レーザー走査型顕微鏡システム
JP2015038539A (ja) * 2012-10-26 2015-02-26 シャープ株式会社 レンズ素子
WO2016185623A1 (fr) * 2015-05-18 2016-11-24 シャープ株式会社 Dispositif de détection de particules fines

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4252447B2 (ja) * 2001-06-22 2009-04-08 カール ツァイス イェナ ゲーエムベーハー 対物レンズ
JP2004219608A (ja) * 2003-01-14 2004-08-05 Kurobane Nikon:Kk 対物レンズ及びこの対物レンズを備える顕微鏡
JP2013152484A (ja) * 2007-07-17 2013-08-08 Olympus Corp レーザー走査型顕微鏡システム
JP2015038539A (ja) * 2012-10-26 2015-02-26 シャープ株式会社 レンズ素子
WO2016185623A1 (fr) * 2015-05-18 2016-11-24 シャープ株式会社 Dispositif de détection de particules fines

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